CAJ | 학술논문

目的分离和培养人脐静脉内皮细胞(HUVECs),对其进行鉴定,并探讨酪氨酸激酶受体-2(Tie-2)基因在HUVECs中的表达情况.方法用胰蛋白酶消化、分离HUVECs并对其进行培养,根据细胞生长特点、形态特征和Ⅷ因子免疫荧光组化技术对细胞进行鉴定.分别采用逆转录-聚合酶链反应(RT-PCR)和SABC免疫细胞化学染色对HUVECs中Tie-2 mRNA和蛋白表达情况进行检测.结果原代培养的HUVECs约于24 h完全贴壁,4～5 d后融合成单层铺路石样结构.Ⅷ因子免疫荧光组化法证实细胞是HUVECs.RT-PCR检测到HuVECs中Tie-2 mRNA条带明显,SABC免疫细胞组化法检测到Tie-2蛋白在HUVECs胞浆、核膜中旱强阳性表达,阳性率为85%以上.结论胰蛋白酶灌流消化法可获得高纯度的HUVECs,Tie-2在HUVECs中旱强阳性表达.
목적 분리화배양인제정맥내피세포(HUVECs),대기진행감정,병탐토락안산격매수체-2(Tie-2)기인재HUVECs중적표체정황.방법 용이단백매소화、분리HUVECs병대기진행배양,근거세포생장특점、형태특정화Ⅷ인자면역형광조화기술대세포진행감정.분별채용역전록-취합매련반응(RT-PCR)화SABC면역세포화학염색대HUVECs중Tie-2 mRNA화단백표체정황진행검측.결과 원대배양적HUVECs약우24 h완전첩벽,4～5 d후융합성단층포로석양결구.Ⅷ인자면역형광조화법증실세포시HUVECs.RT-PCR검측도HuVECs중Tie-2 mRNA조대명현,SABC면역세포조화법검측도Tie-2단백재HUVECs포장、핵막중한강양성표체,양성솔위85%이상.결론 이단백매관류소화법가획득고순도적HUVECs,Tie-2재HUVECs중한강양성표체.
Objective To study the cultural method and identification of human umbilical vein endothelial cells(HUVECs),and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2)in HUVECs.Methods HUVECs were isolated from umbilical veins by the technique of irrigative digestion,and were cultivated in plates.The cells were identified by Ⅷ monoclonal antibody.Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction(RT-PCR)and SABC immuno-cytochemistry.Results HUVECs could adhere to the plates completely after 24 hours,and confluence a monolayer 4-5 days later.The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs.The positive rate was over 85%.Conclusion Highly purified endothelial cells were isolated.And there were overexpression of Tie-2 in HUVECs.